| Objective: To investigate the mechanism of Zhuanggu Zhitong Capsule regulating Notch pathway intervention postmenopausal osteoporosis by observing the difference in expression of Notch1,Jagged1,and HES protein in Notch pathway of ovariectomized rats.Methods: 36 female SD rats were randomly selected,6 of which were removed from the ovary with the same quality of fat as sham operation group,6 were treated as blank group without any treatment,and the remaining 24 rats were prepared with osteoporosis models by bilateral ovariectomy.Divided into model group,RAPA group,3-MA group and Zhuanggu Zhitong capsule group,in total of 4 groups,6 in each group.Dosing started 6 days after surgery for a month.The material was collected on the second day after the last administration,and blood was collected through the abdominal aorta to prepare the corresponding group-containing serum.ELISA was used to determine the content of type I collagen amino terminal elongation peptide(P1NP)in the serum;bone marrow mesenchymal stem cells isolated from the tibia and femur of rats were passed to the second generation;HE staining observation and IHC determination in renal and bone about the expression of HES,Notch1 and Jagged1 protein.Results:(1)HE:Staining found that the trabecular bone spacing of the model group was increased,the trabecular bone cavity was smaller,and the bone collar was incomplete,indicating successful modeling.(2)ELISA:Compared with the blank group,there was no statistically significant difference in the expression of serum P1 NP in the sham operation group(P>0.05).Compared with the sham operation group,the serum P1 NP value of the model group decreased,and there was a significant difference(P<0.01).Compared with the model group,the expression of serum P1 NP in Zhuanggu Zhitong Capsule group,RAPA group and 3-MA group was significantly increased(P<0.01).(3)IHC of bone:The expression of HES,Jagged1 and Notch1 in sham operation group and blank group had no significant difference(P>0.05);compared with model group,the expression of HES,Jagged1 and Notch1 in Zhuanggu Zhitong Capsule group and RAPA group was significantly higher(P<0.01),the expression of Jagged1 and Notch1 in 3-MA group was higher(P<0.01),the expression of HES was lower(P<0.01);Compared with RAPA group,the expression of HES,Notch1 and Jagged1 in 3-MA group decreased significantly(P<0.01).(4)IHC of renal:There wasno significant difference in the expression of Notch1 between the sham operation group and the blank group(P>0.05);compared with the sham operation group,the expression of HES in the blank group was significantly higher(P<0.01),the expression of Jagged1 was significantly lower(P<0.01);Compared with the model group,the expressions of HES,Jagged1 and Notch1 in Zhuanggu Zhitong Capsule group and RAPA group were significantly increased(P<0.01),the expressions of Jagged1 and Notch1 in 3-MA group were significantly decreased(P<0.01),and there was no significant difference in the expression of HES(P>0.05);compared with RAPA group,the expression of HES,Notch1,Jagged1 in 3-MA group was significantly reduced(P<0.01).Conclusion:Zhuanggu Zhitong Capsule can regulate Notch pathway in ovariectomized rats,positively regulate Notch1,Jagged1 and HES protein and interferewith postmenopausal osteoporosis;2.Zhuanggu Zhitong capsules and RAPA can promote the process of bone formation;3.Zhuanggu Zhitong capsules can effectively improve the bone microstructure of PMOP. |
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